Activity-based selections and amplification of non-natural protease substrates

蛋白酶催化肽和蛋白质中酰胺键的水解，在蛋白质合成中发挥重要作用。 大多数生物学过程，并且是许多生物学过程的非常重要的治疗和诊断靶点。 疾病，包括癌症。在这项建议中，开发高效和系统的组合工具 提出了确定前列腺癌中涉及的蛋白酶的底物特异性概况。 这三种蛋白酶的高选择性和特异性底物的产生将有助于设计更多的 选择性诊断工具，以及有可能帮助设计选择性抑制剂。 指导K99阶段：将进行该提案的博士后阶段（具体目标I和II） 在加州大学伯克利分校乔纳森·埃尔曼教授的指导下。K99阶段 目的是开发一种用于产生完全特异性蛋白酶底物的方法。基板 将是肽和基于活性的选择，使用硫酯易裂键将在 DNA结合文库。DMA显示技术将用于编码、路由和放大 图书馆乔恩·埃尔曼是组合化学及其在蛋白酶中的应用领域的领先专家 印刷受体.这一时期将提供组合化学和蛋白酶领域的基本培训 化学.所获得的知识将证明对完成具体目标三和四是非常宝贵的， 成为一名成功的独立科学家。 独立ROO阶段：具体目标III和IV将侧重于技术的应用 具体目标一和二。靶向参与前列腺癌的蛋白酶的肽底物将 作为前列腺癌的前药治疗剂进行评价（特定目标III）。由以下物质提供的基材： 这些实验也将为蛋白酶激活的近红外探针的产生提供基础， 前列腺肿瘤的体内成像。 相关性：该提案有可能通过促进公共卫生和卫生保健， 组合文库的合成和评价（使用基于活性的选择）。拟议研究 开发了提供完全选择性蛋白酶底物的方法。所获得的基材 将用作药物递送剂和选择性成像剂的基础。

Proteases, which catalyze the hydrolysis of amide bonds in peptides and proteins, play essential roles in most biological processes and are very important therapeutic and diagnostic targets for a multitude of diseases, including cancer. In this proposal, the development of efficient and systematic combinatorial tools is proposed to determine the substrate specificity profiles of proteases involved in prostate cancer. Generation of highly selective and specific substrates for these three proteases will aid in designing more selective diagnostic tools as well as have the potential to aid in designing selective inhibitors. Mentored K99 Phase: The postdoctoral phase (Specific Aims I and II) of the proposal will be carried out under the mentorship of Professor Jonathan Ellman at the University of California-Berkeley. The K99 phase goal is to develop a method for the generation of completely specific protease substrates. The substrates will be peptidic and activity-based selections using a thioester scissile bond will be performed on the DNA-bound library. DMA display technology will be utilized for encoding, routing, and amplification of the library. Jon Ellman is a leading expert in the field of combinatorial chemistry and its application to protease substrates. This period will provide essential training in the fields of combinatorial chemistry and protease chemistry. The acquired knowledge will prove invaluable for completion of Specific Aims III and IV and transition into a successful career as an independent scientist. Independent ROO Phase: Specific Aims III and IV will focus on the application of the technology developed in Specific Aims I and II. Peptidic substrates targeting proteases involved in prostate cancer will be evaluated as pro-drug therapeutics for prostate cancer (Specific Aim III). The substrates afforded from these experiments will also provide the basis for the generation of protease-activated near IR probes for the in vivo imaging of prostate tumors. Relevance: This proposal has the potential to greatly improve public health through advancement of the synthesis and evaluation (using activity-based selections) of combinatorial libraries. The proposed research develops methodology that will provide completely selective protease substrates. The substrates obtained will be used as the basis for drug delivery agents and selective imaging agents.